Objective To explore the therapeutic effect of picroside Ⅱ(P Ⅱ)on diabetes nephropathy(DN)rats by regulating fatty acid synthase(Fas)/fatty acid synthase ligand(FasL)signaling pathway.Methods A DN rat model was constructed by combining high sugar and high-fat diet with streptozotocin(STZ)injection.DN rats were grouped into model group(DN group),low,medium and high dose picroside Ⅱ groups(P-L group,P-M group,P-H group),and high dose picroside Ⅱ+Fas recombinant protein group(P-H+rh-Fas group),with healthy rats as control group,and 18 rats in each group.Fasting blood glucose(FBG),body mass,24-hour urinary protein(24h UTP),and renal function(SCr,BUN)were measured in rats.Hematoxylin-eosin(HE)staining and Masson staining were applied to observe the pathological changes in renal tissue of rats;immunohistochemistry and Western blot were applied to detect the expression of RAGE,COL-Ⅳ and pathway proteins,respectively.Results Compared with the control group,rats in DN group showed thickening of the glomerular basement membrane,mesangial proliferation,tubular degeneration,dilation,atrophy,fatty degeneration,obvious collagen deposition,higher levels of FBG,24h UTP,SCr,BUN,TNF-α,IL-6 and IL-1β,body mass loss,and higher expression of RAGE,COL-Ⅳ,Fas and FasL(P＜0.05).Compared with the DN group,the glomerular and tubular lesions were reduced and collagen deposition was decreased in the P-L,P-M and P-H groups,furthermore,the FBG,24h UTP,SCr,BUN,TNF-α,IL-6,IL-1β levels were lower,body mass was higher,and the RAGE,COL-Ⅳ,Fas,FasL expression was lower(P＜0.05).Compared with the P-H group,the renal tissue lesions in the P-H+rh-Fas group worsened,the FBG,24 h UTP,SCr,BUN,TNF-α,IL-6,IL-1β levels were higher,body mass was lower,and the RAGE,COL-Ⅳ,Fas,FasL expression was higher(P＜0.05).Conclusion Picroside Ⅱ exerts therapeutic effects on DN rats by inhibiting the Fas/FasL signaling pathway.

Objective To explore the therapeutic effect of picroside Ⅱ(P Ⅱ)on diabetes nephropathy(DN)rats by regulating fatty acid synthase(Fas)/fatty acid synthase ligand(FasL)signaling pathway.Methods A DN rat model was constructed by combining high sugar and high-fat diet with streptozotocin(STZ)injection.DN rats were grouped into model group(DN group),low,medium and high dose picroside Ⅱ groups(P-L group,P-M group,P-H group),and high dose picroside Ⅱ+Fas recombinant protein group(P-H+rh-Fas group),with healthy rats as control group,and 18 rats in each group.Fasting blood glucose(FBG),body mass,24-hour urinary protein(24h UTP),and renal function(SCr,BUN)were measured in rats.Hematoxylin-eosin(HE)staining and Masson staining were applied to observe the pathological changes in renal tissue of rats;immunohistochemistry and Western blot were applied to detect the expression of RAGE,COL-Ⅳ and pathway proteins,respectively.Results Compared with the control group,rats in DN group showed thickening of the glomerular basement membrane,mesangial proliferation,tubular degeneration,dilation,atrophy,fatty degeneration,obvious collagen deposition,higher levels of FBG,24h UTP,SCr,BUN,TNF-α,IL-6 and IL-1β,body mass loss,and higher expression of RAGE,COL-Ⅳ,Fas and FasL(P＜0.05).Compared with the DN group,the glomerular and tubular lesions were reduced and collagen deposition was decreased in the P-L,P-M and P-H groups,furthermore,the FBG,24h UTP,SCr,BUN,TNF-α,IL-6,IL-1β levels were lower,body mass was higher,and the RAGE,COL-Ⅳ,Fas,FasL expression was lower(P＜0.05).Compared with the P-H group,the renal tissue lesions in the P-H+rh-Fas group worsened,the FBG,24 h UTP,SCr,BUN,TNF-α,IL-6,IL-1β levels were higher,body mass was lower,and the RAGE,COL-Ⅳ,Fas,FasL expression was higher(P＜0.05).Conclusion Picroside Ⅱ exerts therapeutic effects on DN rats by inhibiting the Fas/FasL signaling pathway.

The patient′s ABO blood type and Rh antigen phenotype were identified by monoclonal antibody serum test tube agglutination, and Rh antigen deletion was confirmed by gene sequencing.The ABO blood type and Rh antigen phenotype of the patient were identified using monoclonal antibody serum in vitro agglutination assay, and Rh antigen deletion was confirmed using gene sequencing. The Rh typing saline method showed that the patient was positive for anti D, but negative for anti E, -C, -c, and -e. The saline method for antibody screening showed negative results for cells I to III, positive results for polyamine and anti human globulin tests, positive results for antibody identification cells 1 to 16, and negative results for themselves. Direct anti globulin tests showed negative results. The sequencing results of RhC/E gene showed that exons 9-10 were normal, while exons 1-8 were missing. The patient had a deletion of exons 1-8 of the RhC/E gene, resulting in a loss of Rh antigen E/e and C/c expression. After the first random matching transfusion, the patient produced antibodies targeting E/e and C/c, resulting in an incompatibility between the main and side matching during the second infusion of red blood cell products and the inability to transfuse. In order to solve this situation, first we need to establish a rare blood group bank for Rh C/E gene deletion. Secondly, during the first blood transfusion, a small amount of RH antigen red blood cells should be injected. Stored autologous blood transfusion should also be considered.

Objective To investigate the relationship of blood concentration of tacrolimus with efficacy and safety in treatment of patients with myasthenia gravis.Methods The clinical data of 74 patients with myasthenia gravis admitted in our hospital from January 2013 to December 2015 were analyzed retrospectively.The blood concentrations of tacrolimus were determined by chemiluminescence microparticle immunoassay.Treatment effects were evaluated with clinical relative scoring in China.Results There were 41 men and 33 women in 74 patients with the mean age of (56.7 ± 15.8) years.According to the Osserman classification,there were 7 cases of type Ⅰ,18 cases of type Ⅱ b,27 cases of type Ⅱ a,5 cases of type Ⅲ and 17 cases of type Ⅳ.Forty seven patients underwent thymectomy,including 9 cases of thymoma and 38 cases of hyperplasia.Patients were followed up for (28.1 ± 14.5)days,and the effective rate was 74 ％ (55/74).There was no significant correlation between tacrolimus blood concentration and clinical efficacy.The incidence of adverse reaction of tacrolimus was 22％ (16/74),and the incidence of adverse reactions increased with the increase of blood concentration.Conclusion Tacrolimus is effective in treatment of myasthenia gravis.The correlation of blood concentration of tacrolimus with the clinical efficacy needs to be further studied,but certainly,the monitoring of blood concentration can reduce the incidence of adverse reactions.

Holmes-Adie syndrome (HAS) is a clinical syndrome mainly characterized by tonic pupil and disappearance of tendon reflex. It is mostly idiopathic and can also be seen in cerebral diseases, such as trauma, infection and tumors. However, it is rarely reported to be accompanied with myasthenia gravis (MG). We report a case of MG and HAS, whose clinical manifestations were fluctuation of limb weakness, breathing difficulties, right ptosis. Her pupils were unequal: the left pupil was 3 mm, the right pupil was 2 mm, direct and indirect light reflex was slow in left pupil, and right pupil was sensitive to light reflex. The left eye pupil shrank to 2 mm after dripped pilocarpine diluent for 10 minutes, while the right pupil was still 2 mm. Chest CT examination revealed thymoma. After treatment with thymectomy, glucocorticoid, immunoglobulins and tacrolimus, her symptoms of MG were improved, but the left pupil diameter and light reflex were not changed. Combined with the patient's symptoms, physical signs and examinations, this patient was diagnosed as MG accompanied with HAS.

Objective To investigate the effect of CYP3A5 genetic polymorphism on tacrolimus concentration in myasthenia gravis patients to explore the optimum doses for patients of different genotypes.Methods The CYP3A 5 genotypes of 95 myasthenia gravis patients treated with tacrolimus in our hospital from January 2013 to December 2015,were determined by fluorescence in situ hybridization and tacrolimus concentrations were determined by chemiluminescence microparticle immuno assay.The differences of tacrolimus concentrations/dosage (C/D) ratios,concentration and dosage oftacrolimus were compared among patients of different genotypes.Results There were 14 patients (14.74％) with CYP3A5*l*l,35 (36.84％) with CYP3A5*l*3 and46 (48.42％) with CYP3A5*3*3 in these 95 patients.The allele frequencies of *1 and *3 were 33.16％ and 66.84％,respectively.The C/D ratios of patients from the 3 groups were significantly different (F=24.860,P=0.000):the C/D ratio of CYP3A5*1/*1 patients was significantly lower than that of CYP3A5*1/*3 patients (P＜0.05),and that of CYP3A5*1/*3 carriers was significantly lower than that of CYP3A5*3/*3 patients (P＜0.05).The appropriate doses of tacrolimus of myasthenia gravis patients were different in different genotypes patients:the dose of patients with CYP3A5* 1* 1 was 4-5 mg/d,that of CYP3A5*3*3 was 2-3 mg/d,and that of CYP3A5*1*3 was between that of CYP3A5*1*1 and CYP3A5*3*3.Conclusions The polymorphism of CYP3A 5 genetic polymorphism has effect on plasma concentrations oftacrolimus in patients with myasthenia gravis.Clinical application ofpharmacogenetic studies will be helpful for individualization oftacrolimus appropriate dosage.

Objective To explore the clinical differences between patients with early-onset myasthenia gravis (EOMG)and those with late-onset myasthenia gravis(LOMG).Methods This was a retrospective study enrolling 157 MG patients.Based on the age of onset,patients were divided into the EOMG group(n=85)and the LOMG group(n =72).The groups were compared on clinical characteristics,including clinical manifestations,MG classification,electrophysiological findings on repetitive nerve stimulation(RNS),single fiber electromyography(SFEMG),levels of antibody against acetylcholine receptors(Ach-R Ab),antibody to muscle-specific kinase(MuSK Ab),titin antibody(Titin Ab),ryanodine receptor antibody(RyR Ab),thyroid function,thymectomy,thymus pathology and responses to treatment.Results The mean ages of onset were markedly different [(40.9 ± 9.7) years vs.(62.0 ± 12.2) years,P＜ 0.05] between the EOMG and LOMG groups.The LOMG group was associated with a significantly higher rate of the ocular form(50.0 ％,n=36 vs.32.9％,n=28,P＜0.05),a lower rate of the general form(50.0％,n=36 vs.67.1％,n=57,P＜0.05),and an increased risk of bulbar involvement(41.7％ n=30 vs.23.5％,n=20,P＜0.05)than those in the EOMG group.There was no significant difference in positive rates of RNAS and SFEMG,and levels of AChR Ab,MuSK Ab and double serum negative(DSN)MG between the groups (P＞0.05).Moreover,patients in the EOMG group were more likely to have abnormal thyroid function and higher percentages of receiving steroids,tacrolimus,plasma exchange therapy,and thymectomy (P＜ 0.05).Conclusions The clinical profiles of LOMG are different from those of EOMG in clinical manifestations,thyroid function,thymectomy frequency,striational antibody levels and disease-modifying drug options.

To investigate the effect ofgene down-regulation on early hematopoietic development of zebrafish.Phosphorodiamidate morpholino oligomer (PMO) technology was used to downregulategene expression in Zebrafish. Zebrafish embryos injected phosphorodiamidate morpholino antisense oligonucleotide ofgene mRNA by microinjection at unicellular stage were taken as the experimental group, and those injected meaningless phosphorodiamidate morpholino antisense oligonucleotide were taken as the control. The embryos were collected at 18, 24, 30 and 36 hpf after the fertilization. The real-time fluorescent quantitative PCR (RT-PCR) and whole embryohybridization methods were used to detect the expression of myeloid hematopoietic transcription factorand erythroid hematopoietic transcription factorin zebrafish.RT-PCR showed that the expressions ofanddecreased in the experimental group compared with the control group (all<0.05). Whole embryohybridization showed that the blue-black positive hybridization signals ofandin experimental group were shallow than those in the control group.Myeloid hematopoietic and erythroid hematopoietic of zebrafish are blocked with the downregulation ofgene.
Animals ; Down-Regulation ; genetics ; Embryo, Nonmammalian ; physiopathology ; GATA1 Transcription Factor ; genetics ; metabolism ; Gene Knockdown Techniques ; Hematopoiesis ; In Situ Hybridization ; Lamin Type A ; genetics ; physiology ; Proto-Oncogene Proteins ; genetics ; metabolism ; Trans-Activators ; genetics ; metabolism ; Zebrafish ; embryology ; genetics